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MicroAmpTM Optical 384-Well Reaction Plate

Company: Thermo Fisher Scientific
Catalog#: 4343370
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Quantifying Gene Expression Directly from FACS Using Hydrolysis (TaqMan) Probes on Flash-frozen Cells
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2018-04-20
[Abstract]  The TaqMan Gene Expression Cells-to-CtTM Kit enables reverse transcription directly from cell lysates without the need for isolating RNA. The recommended input range for the lysis reaction is between 10-105 cells, though the upper limit may vary somewhat according to cell type. This protocol is partially adapted from the manufacturer’s protocol (TaqMan Gene Expression Cells-to-CtTM Kit User Guide) for gene expression for use with frozen cell pellets to allow for sample transfer among laboratories and involves experimentally confirmed dilution prior to lysis. Within this constraint, the recommended range for sample cell number for frozen cells is between 40-2 x 105 ... [摘要]  TaqMan基因表达细胞至Ct TM TM试剂盒能够直接从细胞裂解物进行逆转录,而无需分离RNA。裂解反应的推荐输入范围在10-10-5个细胞之间,尽管上限可能根据细胞类型而有所不同。该协议部分适用于制造商的协议( TaqMan Gene Expression Cells-to用于基因表达以便与冷冻细胞团粒一起使用以允许实验室之间的样品转移并涉及在裂解之前通过实验证实的稀释。在这个限制范围内,冷冻细胞的样品细胞数量的推荐范围在40-2×10-5个细胞之间。该试剂盒还设计用于'自制'qPCR主混合物,因为试剂盒中包含的TaqMan Genotyping Master Mix体积可能不足以用于大量样品和/或分析的分析(Smythe和Copren,2008)。细胞计数由荧光激活细胞分选(FACS)提供,并且是该方案的独特方面。【背景】此协议是对TaqMan Gene的修改和实验验证, Expression Cells-to -CT 试剂盒用户指南,出版号为4385117版本E(2012)(Thermo Fisher Scientific / Life ...

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